학술논문

Intracellular flow cytometry complements RT-qPCR detection of circulating SARS-CoV-2 variants of concern
Document Type
article
Source
BioTechniques, Vol 72, Iss 6, Pp 245-254 (2022)
Subject
antiviral drug
duplex RT-qPCR
flow cytometry
nucleocapsid
omicron variant
SARS-CoV-2
Biology (General)
QH301-705.5
Language
English
ISSN
1940-9818
0736-6205
Abstract
Basic and antiviral research on SARS-CoV-2 rely on cellular assays of virus replication in vitro. In addition, accurate detection of virus-infected cells and released virus particles is needed to study virus replication and to profile new candidate antiviral drugs. Here, by flow cytometry, we detect SARS-CoV-2 infection at single cell level and distinguish infected Vero E6 cells from uninfected bystander cells. Furthermore, based on the viral nucleocapsid expression, subpopulations of infected cells that are in an early or late phase of viral replication can be differentiated. Importantly, this flow cytometric technique complements our duplex RT-qPCR detection of viral E and N, and it can be applied to all current SARS-CoV-2 variants of concern, including the highly mutated Omicron variant.